RNF43 p.G659fs 通过 PI3K/AKT/mTOR 信号通路和 HLA-E 上调导致 MSI-high 结直肠癌中 NK 细胞功能障碍
RNF43 p.G659fs leads to natural killer cell dysfunction in MSI-high colorectal cancer through PI3K/AKT/mTOR signaling and HLA-E up-regulation.
CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
英文原题:Large-scale Isolation of Exosomes Derived from NK Cells for Anti-tumor Therapy.
Large-scale Isolation of Exosomes Derived from NK Cells for Anti-tumor Therapy.
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外泌体是脂质双层包裹的囊泡,由细胞主动分泌,含有蛋白质、脂质、核酸等物质,进入靶细胞后具有多种生物学功能。来源于NK细胞的外泌体已被证明具有一定的抗肿瘤作用,并具有作为化疗药物载体的潜在应用。这些进展导致对外泌体的需求很高。尽管外泌体已有大规模工业制备,但仅针对HEK 293T等一般工程化细胞。特定细胞外泌体的大规模制备仍然是实验室研究中的主要问题。因此,在本研究中,我们使用切向流过滤(TFF)浓缩从NK细胞分离的培养上清液,并通过超速离心分离NK细胞来源的外泌体(NK-Exo)。通过对NK-Exo进行一系列表征和功能验证,验证了NK-Exo的表征、表型和抗肿瘤活性。我们的研究为分离NK-Exo提供了一种相当省时省力的方案。
Exosomes are lipid bilayer-enclosed vesicles, actively secreted by cells, containing proteins, lipids, nucleic acids, and other substances with multiple biological functions after entering target cells. Exosomes derived from NK cells have been shown to have certain anti-tumor effects and potential applications as chemotherapy drug carriers.
These developments have resulted in high demand for exosomes. Although there has been large-scale industrial preparation of exosomes, they are only for generally engineered cells such as HEK 293T. The large-scale preparation of specific cellular exosomes is still a major problem in laboratory studies.
Therefore, in this study, we used tangential flow filtration (TFF) to concentrate the culture supernatants isolated from NK cells and isolated NK cell-derived exosomes (NK-Exo) by ultracentrifugation. Through a series of characterization and functional verification of NK-Exo, the characterization, phenotype, and anti-tumor activity of NK-Exo were verified.
Our study provides a considerably time- and labor-saving protocol for the isolation of NK-Exo.
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